title: Gated CW-STED microscopy: a versatile tool for biological nanometer scale investigation.
abstract: Stimulation emission depletion (STED) microscopy breaks the spatial resolution limit of conventional light microscopy while retaining its major advantages, such as working under physiological conditions. These properties make STED microscopy a perfect tool for investigating dynamic sub-cellular processes in living organisms. However, up to now, the massive dissemination of STED microscopy has been hindered by the complexity and cost of its implementation. Gated CW-STED (gCW-STED) substantially helps solve this problem without sacrificing spatial resolution. Here, we describe a versatile gCW-STED microscope able to speedily image the specimen, at a resolution below 50 nm, with light intensities comparable to the more complicated all-pulsed STED system. We show this ability on calibration samples as well as on biological samples.
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy. Electronic address: giuseppe.vicidomini@iit.it.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vicidomini
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy; Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genoa, Italy.
authors.forename: Ivan Coto
authors.initials: IC
authors.lastname: Hernández
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy.
authors.forename: Marta
authors.initials: M
authors.lastname: d'Amora
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy.
authors.forename: Francesca Cella
authors.initials: FC
authors.lastname: Zanacchi
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Bianchini
authors.affiliation: Department of Nanophysics, Istituto Italiano di Tecnologia, Via Morego 30, 16163 Genoa, Italy; Department of Physics, University of Genoa, Via Dodecaneso 33, 16146 Genoa, Italy. Electronic address: alberto.diaspro@iit.it.
authors.forename: Alberto
authors.initials: A
authors.lastname: Diaspro
mesh_terms: D000818:Animals; D019556:COS Cells; D002138:Calibration; D002522:Chlorocebus aethiops; D005456:Fluorescent Dyes / Q000737:chemistry; D006367:HeLa Cells; D006801:Humans; D057230:Limit of Detection; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods; D008870:Microtubules / Q000648:ultrastructure*; D049329:Nanostructures / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes
keywords: Spatial resolution; Stimulated emission depletion microscopy; Super-resolution microscopy